Abstract
Charybdis lucifera, a seafood delicacy in many parts of the world, are discarded from commercial fishery operations of Karnataka coast (India), since it is not used for consumption in the region. Globally, in light of reports on overfishing and reduction in fishery, reducing the fishing pressure on many conventional species are becoming a priority.
To meet the probable reduction in availability of seafood, due to the regulations on the fishery of conventional species, there is need to bring in more non-conventional species to commercial status.
Information on nutritional quality of the non-conventional resources is the essential pre-requisite for effective use of these resources. This study, therefore, was focused to promote a non-conventional crab resource to commercial scale by unveiling information on their abundance, nutritional quality and economic importance. Our study revealed that C. lucifera is a regular component of trawl bycatch of Karnataka coast and annual average landing during 2015-2016 was estimated at 45 t. The species was found to have high meat content (27.2 ±5.9%) which is comparable or even better than most of the conventional edible crabs. Crude protein in male and female were 80.1 and 79.0% respectively, with high nutritive essential amino acid profile. Moreover, the species was found to possess rich PUFA, Eicosapentaenoic acid (13.5%) and Docosahexaenoic acid (16.8%) content, which qualifies it as a high quality seafood. At present these crabs are being disposed as “low value bycatch” for fertilizer and
poultry feed after drying, with other trash fishes realizing an average price of Rs.5/kg which can be increased at least ten times, once it is popularized as a food crab. Moreover, its commercial use could eventually lead to increase in the landings of the species by discards.
Even with the present annual catch data and with the projected value escalation, the revenue could be increased up to Rs.2.3 million (approx.30,000 $) every year, which definitely will help in improving the livelihood status of fishermen involved in coastal fishery. Its potential as a source of bio-active components like chitin, chitosan and astaxanthin was also projected in the study to supplement its economic significance.
Keywords: Non-conventional resources, trawl discard, Chraybdis lucifera, nutritional quality, southwest coast of India, livelihood improvement
Introduction
Globally, since 2013, one in eight people are deprived of enough food and most of them are in extreme hunger (FAO, IFAD and WFP, 2013). To feed a population, which was projected to reach nine billion by 2050 (Duarte et al., 2009), the living resources
Investigations on the non-
conventional crab fishery resource, Charybdis lucifera (Fabricius, 1798) from Karnataka coast (India) and the need to commercialize the resource
K. Yogesh Kumar, A. P. Dineshbabu* and Sujitha Thomas Research Centre of ICAR- CMFRI, Mangalore, Karnataka– 575 001, India.
*Correspondence e-mail: dineshbabuap@yahoo.co.in
Received : 05 Oct 2020 Accepted : 03 May 2021 Published : 15 May 2021
Original Article
from marine and coastal zones (comprising of 70% of the globe) are considered as potential source of healthy diet (Walford and Wilber, 1955; Duarte et al., 2009; Thilsted et al., 2014). Over the years due to adoption of technological advancements in fishing, fish production increased substantially. The production from the wild was 81.2 million t in 2015, wild catches were reported to be approaching its sustainable limit, as almost 90% of the global fish stocks are overfished (FAO, 2018).
Conservation and management of the conventional resources and utilization of non-conventional fishery resources are considered viable options to meet the growing demand for fish from wild (Sajeevan and Nair, 2006). As far as exploitation and utilization of non-conventional marine resources are concerned, most of the earlier workers focused on exploitation and utilization of deep sea varieties from distant waters (Suda, 1973), however there were several difficulties in developing sustainable fishery for this resources, for example: technical difficulties, economic loss in deep sea fishing operation. Low resilience capacity of deep sea species, also made commercial deep-sea fisheries unsustainable (Elliott et al., 2012). Further it was estimated with
“state of art” technological support, deep sea fishery could only contribute less than 1% of sea food. In this context, bringing more non-conventional fish protein to human diet is projected as one of the few alternatives to provide affordable protein supplement to growing population (Thilsted et al., 2014). Many non–conventional species with better preservation techniques and knowledge on the nutritional quality were brought out to provide human nutrition (Lai and Leung, 2003; Ilavarasan et al., 2015a, 2015b). Apart from utilizing for direct human consumption, guidelines are also available for preparation of food additives from conventional and non-conventional species (Mohanty and Roy, 1955; Olden, 1960). In addition to nutritional security, by using fishes from “discards” for human consumption, the economic status of the fishermen could be improved considerably with higher value realized for their catch (Thilsted et al., 2014).
In the present study attempts are made to explore the possibilities of using a species discarded during fishing as human nutritional supplement. While considering a non-conventional species for commercial exploitation, there are some prerequisites to be followed, to popularise its exploitation and consumption.
Ascertaining the taxonomic status of the species, awareness of its commercial use elsewhere, ascertaining its availability in space and time for commercial exploitation are some of the prerequisites involved in the process. Even though availability of the species along Karnataka coast is known (Dineshbabu et al., 2011), to promote commercialisation of the species several aspects of biology, distribution and nutritional quality should be explored. Kumar et al. (2019) made a preliminary study on distribution and abundance of this species.
Most importantly, while considering a species for popularisation, information on its meat content, its nutritional status and value addition are of prime importance. Preliminary studies on nutritional quality of C. lucifera was carried out on the east coast which suggests the possibility of acceptance of the species for human consumption (Ilavarasan et al., 2015a; Kumari et al., 2015; Ramamoorthy et al., 2016). It was also suggested that there is regional variation in nutritional quality of this species, and in this context, the objective of the present study was to evaluate the distribution, abundance and nutritional quality of C. lucifera from west coast of India.
Meat content, proximate composition, mineral content, amino acid profiles and fatty acid profiles were carried out in the present study, to explore its economic importance and also the chitin, chitosan and astoxanthin content available in the species were studied in detail.
Material and methods Study area and sampling
Investigation was carried out from single day trawlers operating within 50 m depth along Karnataka coast (between 12.50N 750E and 150N and 74.50E) south west coast of India (Fig.1).
Study was carried out for two years from January 2016 to December 2017. A part of C. lucifera catch was kept in ice
Fig. 1. Study area of C. lucifera.
packing and brought to the laboratory. In the laboratory, crabs were sex-wise segregated; measurements (carapace width to nearest mm and weight to nearest g) were noted. For Identification of C. lucifera, the keys provided by Mizzan and Vianello (2009) was followed.
Meat content
For extracting meat, carapace of individual crab was removed and meat content, including the meat from chelate legs, were removed. The meat was weighed and transferred to petri dish for further analysis. The male crabs used for the study ranged from 35 to 80 mm carapace width, weighing 18 to 108 g. In males 79 crabs having hard shells (inter-moult) were used for the study. In females from the inter-moult crabs 28 crabs without berried eggs were selected for the study. From the females 28 females 46 to 63 mm (CW) weighing 15 to 36 g. The overall meat percentage, meat percentage of males and females and meat percentage of the crabs in different size groups (carapace width) were derived (Balasubramanian and Suseelan, 2001).
Proximate analysis
Moisture, total protein, total lipids, ash and total carbohydrate of male and female crab were evaluated based on standard methods.
Crude protein (N×6.25) was evaluated by micro-Kjeldahl method (AOAC, 2012). The quantity of carbohydrate was determined by Phenol Sulphuric acid method (Dubois et al., 1956). All determinations were done in triplicates. Mineral estimation was carried out by the method of inductively coupled plasma-optical emission spectrometric (ICP-OES) determination of elements using microwave-assisted digestion (Horwitz and Latimer, 2005).
The amino acid content of crab meat was determined based on the study of Hofmann et al. (2003) using isotope with gas chromatography–combustion–isotope ratio mass spectrometry (GCC-IRMS/MS) (GC: Hewlett-Packard 58590 series II, Germany;
combustion series II-interface, IRMS MAT 252, Finnigan MAT, Germany; MS: GCQ, Finnigan MAT, Germany). The capillary column of dimension 50 m × 0.32 mm id. × 0.5 µm BPX5 (SGE) was connected to gas chromatography. The flow of carrier gas (helium) was maintained at 1.5 ml/min, with the head pressure 13 psi. The details of temperature program are given in Table 1. Classification of Nelson and Cox (2004) was adopted for classification of amino acids as nutritionally “essential” or
“nonessential” or “conditionally essential”.
Hot extracted crude fat from crab meat was used for determining the fatty acid methyl esters (FAMEs). The method suggested by Padua-Resurreccion and Banzon (1979) was used for this process.
The analytical techniques such as FAMEs were quantified by gas chromatographer (GC2010, Shimadzu, Japan) equipped with
fused silica column (BPX-70) and flame ionization detector (FID).
The identification of peaks obtained from the lipid profiling was determined by comparing with National Institute of Standards and Technology Library (NIST 11 mass spectrometry library;
NIST/EPA/NIH; version #2011). Further analysis was carried out following the methodology suggested by Nareshkumar (2007).
Chitin was extracted by demineralization and de-proteinization methods (Liu et al., 2012) and for chemical composition followed (AOAC, 2012) methodology. Five gram of extracted Chitin was then subjected to deacetylation according to the method of Anand et al. (2014) and Chemical composition estimated by using standard method (AOAC, 2012). The surface morphological appearance of chitin and chitosan were examined with SEM (JSM 6380LA; JEOL, Tokyo, Japan). The presence of chitin and chitosan were confirmed by infrared spectrophotometer (Shimadzu FTIR-8700), characterized from 500 to 4,000 cm-1. Astaxanthin extraction was done by following the method of Dalei and Sahoo (2015). Quantification of astaxanthin was done by using the method of Kelley and Harmon (1972).
The presence of astaxanthin was confirmed by infrared spectrophotometer (Shimadzu FTIR-8700), characterized from 500 to 4,000 cm-1.
Results and discussion Species description
Species identification was done using standard identification keys (Nguyen, 2002).
Classification Phylum: Arthropoda Subphylum: Crustacea Order: Decapoda Family: Portunidae Genus: Charybdis
Species: Charybdis (Charybdis) lucifera (Fabricius, 1798) (Fig. 2)
Table 1. Temperature program for GC-C-IRMS/MS of Amino acid analysis
Time (min) Temperature (°C) Temperature/min
1 50 Start
10 50-100 10°C/min
10 100-175 3°C/min
10 175-200 3°C/min
10 250 stop
C. lucifera is an edible variety which is commercially exploited and marketed along east coast of India (Ilavarasan et al., 2015a). Along west coast of India this species is not considered for human consumption and often discarded or used in fish meal plants. Studies on the distribution of the species showed that they have been a part of the bycatch of single day trawl throughout the trawling season and is extensively distributed all along Southwest coast of India (Kumar et al., 2019) and annual average landing in Karnataka coast during 2015-2016 was estimated at 45 t which is higher than the crab landings from east coast of India (Ilavarasan et al., 2015a).
Meat content
Studies showed that overall meat content for the species was 27.2 ±5.90%. In males the average meat content was 27.7 ±5.71% and in females it was 22.1±5.78%. Size group- wise meat content in males and females are given in the Table 2.
It is an established fact that male crabs yield more meat both from body and claws when compared to females (Sreelakshmi et al., 2016). Even though the meat content in C. lucifera is not high as in Scylla tranquebarica (32.99%) reported by Sreelakshmi et al.
(2016) and as in blue swimmer crab Portunus pelagicus (32%) (Wu et al., 2010), but is comparable with those in S. serrata (30.44%) reported by Sreelakshmi et al. (2016) and was higher than in Chinese mitten crab Eriocheir sinensis (24.2%) as stated by Chen et al. (2007), the deep water crab Charybdis smithii (15.3%) (Balasubramanian and Suseelan, 2001), Atlantic spider crab Maja brachydactyla (17%) (Marques et al., 2010) and brown crab Cancer pagurus (23%) (Barrento et al., 2010). The present study throws light on the huge potential of crab meat availability if exploited commercially.
Biochemical compositions
Present study was conducted on dry weight basis. The moisture content in males and females were 89.27 and 84.31% respectively (Table 3), Crude protein formed 80.10 and 79.01% in males and females respectively. Apart from their delicacy, marine crustacean resources are well known for richness in nutrition (Heu et al., 2003). Considering the health benefits of these sea food, there are growing number of research for promoting crustacean consumption (Rosa and Nunes, 2003; Chen et al., 2007). Crabs specially crab muscle with their high protein and low fat and cholesterol content is getting acceptability as a healthy food globally (Barrento et al., 2010). Studies conducted by Ramamoorthy et al. (2016) showed that protein content in C. lucifera was higher (22.57%) than conventional species P. pelagicus (20.15%). In the present study, carbohydrate in C. lucifera was 3.68% in males and 5.61% in females (Table 1). Ramamoorthy et al. (2015) found higher percentage of carbohydrate content in C. lucifera (1.17%) when compared to the conventional species P. pelagicus (0.54%), whereas the lipid content was comparatively lower than in C. lucifera compared to P. pelagicus (2.15 %). Since the present study was conducted on dry weight basis, direct comparison to the values observed by Ramamoorthy et al. (2016) was not possible. However comparison with studies conducted by Lyla et al. (2017) supports the nutritional superiority of C. lucifera over conventional species P. pelagicus and P. sanguinolentus Fig. 2. C. lucifera (dorsal view)
Table 2. Meat content analysis of C. lucifera male and female. n=3, means ± SD Classification for meat content Average meat weight
(percentage) Overall meat content for the species 27.2±5.90
Meat content in males 27.7±5.71
Males, CW range 30-40mm 20.0±0.42
Males, CW range 40-50mm 26.5±3.88
Males, CW range 50-60mm 28.1±5.82
Males, CW range 60-70mm 27.2±5.98
Males, CW range 70-80mm 29.3±6.19
Meat content in females 22.1±5.78
Females, CW range 30-40mm 21.7±6.18
Females, CW range 40-50mm 26.3±0.97
Values are given as mean ± SD from triplicate determinations.
Table 3. Proximate composition of C. lucifera male and female (%, dry weight) n=3, means ± SD
Parameters (%) C.lucifera male C.lucifera female
Dry matter 10.73±0.22 15.69±0.41
Moisture 89.27±0.21 84.31±0.27
Crude Protein 80.10±0.09 79.01±0.07
Crude fat 3.00±0.00 4.92±0.18
Crude ash 13.22±0.02 10.46±0.21
Carbohydrate 3.68±0.04 5.61±0.08
(Table 4) as these two crabs were having 73.71 and 73.97% of protein. The protein content found in C. lucifera in the present study was higher than those reported in Chinese mitten crab (E. sinensis) (Chen et al., 2007), Podopthalmus vigil (Sudhakar et al., 2011) and Portunus sanguinolentus (Sudhakar et al., 2009).
The richness of amino acids, which are the building blocks of proteins also determines the supremacy of seafood. In east coast of India Ilavarasan et al. (2015a) observed that the species to have vast potential in pharmaceutical formulation, with identification of 9 essential amino acids from the species, whereas present study showed that the species along west coast of India has 9 non-essential and 8 essential amino acids (Table 5) Ramamoorthy et al. (2016) while conducting a comparative
study of the species recorded higher amount of amino acids in C. lucifera (3.92 g/100g) compared to P. pelagicus (2.79 g/100g).
Richness of amino acids in C. lucifera from west coast of India is presented in Table 5. Total essential amino acid content of C. lucifera was above 42.30% which is considered as very good for human nutrition.
Marine lipids with long-chain polyunsaturated fatty acids has proved to have cardio protective action (Kris-Etherton et al., 2002; Ramamurthy et al., 2015). In the present study, C. lucifera is found to have low fat content (3% in male and 4.92% in female), which was lower than those reported by Lyla et al. (2017) in P. pelagicus and P. sanguinolentus, which were 5.68 and 6.96% respectively (Table 3). The sea food with low fat and rich long chain polyunsaturated fatty is reported to have high dietary significance in human growth (Shahidi and Wanasundara, 1998, Dunstan et al., 2007; Su et al., 2008) and C. lucifera found to be rich in PUFA, Eicosapentaenoic acid and Docosahexaenoic acid content with 13.54 and 16.80%
respectively (Table 6), similar findings of comparatively high percentage of PUFA in C. lucifera was reported in east coast also (Ramamoorthy et al., 2015). Mineral analysis of crab meat showed 8 minerals, Sodium, Potassium, Magnesium, Calcium, Manganese, Iron, Zinc and Copper in the present study (Table 7) is one more from those reported from east coast (Kumari et al., 2015) in which copper was absent.
Bioactive compounds
Crustaceans are considered as a good source of Chitin (Kaur and Dhillon, 2013) which is having many nutritional and medicinal utility. It was estimated that approximately half of weight of major crustaceans are formed of chitin (Islam et al., 2004). C. lucifera is a rich source of chitin and the analysis of exoskeleton showed that it contained 16.72% protein, (Table 8). Percentage of extractable protein from the shell was 12.23%
which is comparable with the chitin from commercial shrimps (12.46%) (Table 9). The presence of chitosan is confirmed by FTIR analysis compared with the study literature on crab by Quimque and Acas (2015). FTIR of extracted shell Chitosan of C. lucifera was found to be 3348 cm for hydroxyl group (-OH). Astaxanthin is a xanthophyll carotenoid which is found in various microorganisms and marine animals, which has
Table 4. Proximate components of selected C. lucifera in comparison with conventional commercial species from the coast (%, dry weight of males) n=3, means ± SD
Parameters (%) C. lucifera P. pelagicus P. sanguinolentus
Moisture 89.27±0.21 80.92±0.8 81.61±0.6
Protein 80.10±0.09 73.71±0.1 73.97±0.1
Lipid 3.00±0.00 5.68±0.01 6.96±0.1
Carbohydrate 3.68±0.04 1.39±0.8 1.40±0.5
Ash 13.02±0.02 12.06±0.6 12.66±0.0
Lyla et al. (2017)
Table 5. Amino acid composition of C. lucifera, male and female (%, dry weight) n=3, means ± SD
Amino acid (g/100g) C.lucifera male C.lucifera female Non essential
Asp 4.79±0.09 4.26±0.09
Glu 9.05±0.09 8.85±0.02
Ser 2.34±0.01 2.42±0.01
Gly 14.64±0.05 13.46±0.12
Arg 8.02±0.06 8.91±0.07
Ala 8.70±0.02 8.47±0.03
Pro 5.79±0.09 7.28±0.09
Tyr 2.72±0.07 3.85±0.01
Cys 0.31±0.01 0.29±0.00
Essential
Ile 4.81±0.07 4.86±0.11
Leu 7.54±0.05 7.46±0.01
Phe 3.69±0.09 3.77±0.03
Lys 13.31±0.19 12.53±0.10
His 2.04±0.01 1.64±0.03
Thr 3.15±0.02 3.03±0.08
Val 5.32±0.01 5.27±0.07
Met 2.44±0.03 2.37±0.07
TEAA 42.30±0.47 40.93±0.50
TAA 98.66±0.96 98.72±0.94
Classification of AAs as nutritionally “essential” or “nonessential” or “conditionally essential” is as per Nelson and Cox (2004).
great demand in food, feed, nutraceutical and pharmaceutical applications. Suganya and Asheeba (2015) and Sachindra et al.
(2005) extracted astaxanthin from Portunus sanguinolentus, Callinectes sapidus and Paralithodes brevipes, Charybdis feriata and the present study showed that C. lucifera also can serve as a source for astaxanthin with extractable astaxanthin of 9.19 µg /g shell wet weight.
SEM pattern figure represents the SEM photographs of commercial shrimp chitin (Fig. 3) and the chitin obtained from the exoskeleton of C. lucifera, exhibited dense and firm surface
Table 6. Fatty acid methyl esters (g/100g lipid) of C. lucifera male and female (dry weight) n=3, means ± SD
Fatty acids (g/100g) Charybdis lucifera (male) Charybdis lucifera (female)
Saturated fatty acid
Palmitic acid C16:0 24.75±0.21 25.05±0.03
Heptadecanoic acid C17:0 3.32±0.07 3.02±0.01
Stearic acid C18:0 18.89±0.27 19.13±0.11
Hexanoic acid C6:0 0.36±0.06 0.29±0.09
Octanoic acid C8:0 0.65±0.12 0.99±0.02
Myristic acid C14:0 1.00±0.01 1.43±0.12
Behenic acid C22:0 0 0
Pentadecanoic acid C15:0 0 0
tricosanoic acid C23:0 0 0
Lignoceric acid C24:0 0 0
Arachidic acid C20:0 0 0
Monounsaturated fatty acid
Palmitoleic acid C16: 1 Cis 2.67±0.07 2.73±0.13
Oleic acid C18 : 1 Cis 15.68±0.18 13.47±0.23
Eicosenoic acid C20:1 Cis 0 0
Polyunsaturated fatty acid
alfa Linolenic acid C18: 3 Cis 0 0
Linoleic acid C18:2 Cis 2.34±0.26 3.10±0.06
Eicosapentaenoic acid C20 : 5 Cis 13.54±0.32 14.79±0.41
Docosahexaenoic acid SFA
MUFA PUFA PUFA/SFA
C22: 6
16.80±0.03 48.97±0.74 18.35±0.25 32.68±0.61 0.66±0.82
16.00±0.00 49.91±0.38 16.20±0.36 33.89±0.47 0.67±1.23
Table 7. Mineral content of C. lucifera male and female (dry weight) n=3, means ± SD
Dry weight mg/100gm C. lucifera male C. lucifera female
Copper 0.95±0.36 0.65±0.36
Zinc 3.94±0.63 3.81±1.02
Manganese 0.07±0.03 0.07±0.05
Iron 2.48±0.45 1.80±0.27
Magnesium 69.40±0.32 48.80±0.28
Sodium 651.80±1.01 738.30±0.53
Potassium 251.80±0.51 254.20±0.42
Calcium 113.56±0.32 320.80±0.82
Table 8. Chemical composition of shell (exoskeleton) of crab C. lucliera (dry weight) n=3, means ± SD
% Charybdis lucifera
Shell moisture 64.02± 0.30
Protein 16.72± 0.22
fat 0.72± 0.02
Ash 47.21± 0.15
Table 9. Chemical composition of Chitin from the shell of C. lucliera and commercial shrimp Chitin (dry weight) n=3, means ± SD
Chitin (%) Commercial shrimp
chitin Charybdis lucifera
Yield - 10.4± 0.09
Moisture 7.28 ± 0.16 8.12± 0.02
Protein 12.46 ± 0.33 12.23± 0.20
fat 0.54 ± 0.02 0.00
Ash 0.09 ± 00 0.0981± 0.01
morphology images under the electron microscopic examination at 100X to 2000X magnification (Fig. 4). The surface morphology of the chitin of C. lucifera at 100X and 1000X exhibited clear nanofibres. SEM photographs of commercial shrimp chitosan (Fig. 5) and the chitosan from C. lucifera exhibited dense and firm surface morphology images under the electron microscopic examination at 100X to 1000X magnification (Fig. 6). The surface morphology of chitosan from C. lucifera at 100X and 1000X exhibited clear nanofibres. The porous structure of chitin is used in metal ion absorption and tissue engineering and the fibrillary structure can be used in textiles (Zelencova et al., 2015). There are several commercial brands already available in the global markets (Sastry et al., 2015).
Present study showed that C. lucifera is rich in nutrients, is a good source of high quality chitin, chitosan and astaxanthin.
At present the species is disposed as low value bycatch for fertilizer and poultry feed after drying, along with other trash fishes realizing an average price of Rs.5/kg. Often the species is discarded at sea, due to lack of facility for drying in most of
the landing centres. The meat content in these crabs are as high as 27% which is comparable with most of the edible species in India, denoting the potential of these crabs for domestic consumption as well as export. Once it is popularized as human food with efficient processing and marketing methods, increased price can be obtained, which will be a great boon for the fishermen involved in the coastal fishery. If Rs. 50/kg is earned for the catch, with the annual landing reported. (45t in 2015-2016), the total value realized for the catch can escalate to Rs.2.3 million (approx.30,000 $), eventually improving the livelihood status of the fishermen.
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